Autoinjector comprising high concentration formulation of vedolizumab

This application claims the benefit of U.S. Provisional Application 62/802,671 filed on Feb. 7, 2019. The entire contents of the foregoing application is hereby incorporated by reference.

The instant application contains a Sequence Listing which has been submitted in ASCII format via EFS-Web and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Feb. 5, 2020, is named T103022_1080US1_Sequence_Listing.TXT and is 4,096 bytes in size.

Advances in biotechnology have made it possible to produce antibodies for pharmaceutical applications using recombinant DNA techniques. Because antibodies are larger and more complex than traditional organic and inorganic drugs (i.e., possessing multiple functional groups in addition to complex three-dimensional structures), the formulation of such antibodies poses special problems. For a therapeutic antibody to remain biologically active, a formulation must preserve the conformational integrity of at least a core sequence of the protein's amino acids, while at the same time protecting the protein's multiple functional groups from degradation.

Antibodies have a large number of formulation options, including the state of the formulation, e.g., lyophilized, spray-dried, liquid, and not one approach or system is suitable for all proteins. Several factors to be considered have been reported and are known in the art, including aggregation levels of the antibody in a formulation (See e.g., Wang, et al., J. Pharm Sci. 96:1-26 (2007)).

In addition to identifying a formulation in which the therapeutic antibody will remain stable, delivery of the formulation in a safe and effective way to the patient is critical. Effectively combining suitable delivery of the therapeutic antibody to the patient with a stable formulation can be challenging.

Vedolizumab is a relatively hydrophobic antibody. Coupled with the dose amounts needed for therapeutic efficacy, formulating vedolizumab is particularly challenging, especially when administered subcutaneously.

The invention relates, at least in part, to liquid pharmaceutical formulations having a high concentration of an anti-alpha4beta7 antibody, e.g., 90 mg/ml or greater, that are suitable for subcutaneous delivery to a human patient.

Liquid pharmaceutical formulations described herein provide stability as a liquid formulation which is not dependent on lyophilization, and have relatively low aggregates making them suitable for pharmaceutical use. In one embodiment, stability and safety, e.g., low level of aggregate, are coupled with a viscosity suitable for subcutaneous delivery, e.g., using an autoinjector like those described herein.

In one embodiment, the liquid pharmaceutical formulation comprises at least about 90 mg/ml of the anti-alpha4beta7 antibody; at least about 100 mg/ml of the anti-alpha4beta7 antibody; at least about 110 mg/ml of the anti-alpha4beta7 antibody; at least about 120 mg/ml of the anti-alpha4beta7 antibody; at least about 130 mg/ml of the anti-alpha4beta7 antibody; at least about 140 mg/ml of the anti-alpha4beta7 antibody; at least about 150 mg/ml of the anti-alpha4beta7 antibody; or at least about 160 mg/ml of the anti-alpha4beta7 antibody. In some aspects, the formulation comprises a concentration of the anti-alpha4beta7 antibody between about 90 mg/ml and 200 mg/ml, between about 90 mg/ml to 120 mg/ml, between about 150 mg/ml to 200 mg/ml, between about 50 mg/ml and 180 mg/ml, between about 60 mg/ml and 170 mg/ml, about 70 mg/ml and 160 mg/ml, about 100 mg/ml and 160 mg/ml, or about 108 mg/ml. In one aspect the liquid pharmaceutical formulation has an antibody concentration of about 155 mg/ml to 180 mg/ml. In some embodiments, the liquid pharmaceutical formulation contains at least about 90 mg/ml of anti-alpha4beta7 antibody; at least about 100 mg/ml of anti-alpha4beta7 antibody; at least about 105 mg/ml anti-alpha4beta7 antibody. In one embodiment, the liquid pharmaceutical formulation contains about 90 mg/ml to about 240 mg/ml of anti-alpha4beta7 antibody. In one embodiment, the liquid pharmaceutical formulation contains about 90 mg/ml to about 120 mg/ml of anti-alpha4beta7 antibody. The formulation can contain at least about 160 mg/ml anti-alpha4beta7 antibody. The formulation can contain about 150 to about 200 mg/ml antibody or about 165 mg/ml antibody.

In some aspects, the liquid pharmaceutical formulation comprises a 108 mg dose of the anti-alpha4beta7 antibody. In one embodiment, the liquid pharmaceutical formulation comprises about 54 and 300 mg, about 100 and 216 mg, about 100 and 300 mg, about 95 to 120 mg, about 200 to 230 mg, about 100 to 200 mg, or about 108 mg of the anti-alpha4beta7 antibody.

In certain embodiments, the liquid pharmaceutical formulation comprises at least 90 mg/ml of anti-alpha4beta7 antibody, a buffer, and an excipient in an amount that results in the formulation having a viscosity suitable for subcutaneous delivery via prefilled syringe, an autoinjector, or other device suitable for self-administration.

In certain embodiments, the liquid pharmaceutical formulation can also contain a surfactant, a polysorbate (e.g. polysorbate 20, polysorbate 80), a poloxamer, or any combination thereof.

The liquid pharmaceutical formulation can have a pH between about 5.5 and about 7.5. In certain embodiments, the liquid pharmaceutical formulation has a pH between about 6.0 and about 7.0. In certain embodiments, the liquid pharmaceutical formulation has a pH between about 6.2 and about 7.0. The pH of the liquid pharmaceutical formulation can be between about 6.5 and about 6.8. In certain embodiments, the liquid pharmaceutical formulation can have a pH between about 6.1 and about 7.0, or between about 6.2 and 6.8. In certain embodiments, the liquid pharmaceutical formulation has a pH of about 6.3 to 6.7.

In another aspect, the liquid pharmaceutical formulation can be used to deliver an alpha4beta7 antibody for treating an inflammatory bowel disease in a human subject. In certain embodiments, administration is subcutaneous. The administering can be self-administering. The inflammatory bowel disease can be Crohn's disease or ulcerative colitis. The inflammatory bowel disease can be moderate to severely active ulcerative colitis. The treatment can result in mucosal healing in patients suffering from moderate to severely active ulcerative colitis. The patient may have previously received treatment with at least one corticosteroid for the inflammatory bowel disease. The patient may concurrently receive treatment with at least one corticosteroid for the inflammatory bowel disease. The dosing regimen can result in a reduction, elimination or reduction and elimination of corticosteroid use by the patient.

In some embodiments, the stable liquid pharmaceutical formulation has about 1.0% or less aggregate formation after 1 month at 40 degrees Celsius. In some embodiments, the stable liquid pharmaceutical formulation has about 0.9% or less aggregate formation after 1 month at 40 degrees Celsius. In some embodiments, the stable liquid pharmaceutical formulation has about 0.8% or less aggregate formation after 1 month at 40 degrees Celsius. In some embodiments, the stable liquid pharmaceutical formulation has about 0.7% or less aggregate formation after 1 month at 40 degrees Celsius. In some embodiments, the stable liquid pharmaceutical formulation has about 0.6% or less aggregate formation after 1 month at 40 degrees Celsius. In some embodiments, the stable liquid pharmaceutical formulation has about 0.5% or less aggregate formation after 1 month at 40 degrees Celsius. In some embodiments, the stable liquid pharmaceutical formulation has about 0.4% or less aggregate formation after 1 month at 40 degrees Celsius. Aggregation can be determined according to standard SEC methods known in the art.

A liquid pharmaceutical formulation described herein has a viscosity suitable for delivery in an autoinjector, e.g., 2 to 70 cP. In one embodiment, the volume of the liquid pharmaceutical formulation in the autoinjector can be about 1 mL or less, about 0.9 mL or less, about 0.8 mL or less, about 0.6 mL or less, about 0.5 mL or less, or about 0.4 mL or less, and comprises a dose of about 100 to 110 mg of a humanized anti-alpha4beta7 antibody.

In yet another aspect, the invention relates to an article of manufacture, comprising a container, a liquid pharmaceutical formulation described herein, and instructions for its use.

In one embodiment of the invention, an autoinjector described herein is configured to deliver about 0.5 mL to about 1 mL of a formulation described herein, wherein the autoinjector comprises a tubular body portion; a syringe containing a humanized anti-alpha4beta7 antibody and including a plunger rod; a syringe holder configured to receive the syringe; and a resilient member configured to apply a force on the plunger rod to expel the medicament from the syringe when the autoinjector is activated. In one embodiment, the resilient member is a tension spring or a coil spring.

In one embodiment, the resilient member is configured to apply a force on the plunger rod between about 2-20 Newtons (N), about 3-17 N, about 4-15 N, about 7-13 N, about 5-10 N, about 5-8 N, or about 10N.

In one embodiment, the force applied by the resilient member is based, at least in part, on the viscosity of the formulation being delivered.

The invention relates to a pharmaceutical formulation comprising an anti-alpha4beta7 antibody suitable for delivery using a prefilled syringe.

The term “pharmaceutical formulation” refers to a stable composition containing an active ingredient, such as a therapeutic antibody, in such form as to permit the biological activity of the active ingredient to be effective, and which contains no additional components which are significantly toxic to a subject to which the formulation would be administered. A pharmaceutical formulation is suitable for administration, e.g., via subcutaneous delivery, to a human subject and includes only pharmaceutically acceptable excipients, diluents, and/or other additives deemed safe by the Federal Drug Administration or other foreign national authorities. Pharmaceutical formulations include liquid, e.g., aqueous, solutions that can be directly administered.

A “stable” antibody formulation is one in which the antibody therein substantially retains its physical stability and/or its chemical stability and/or its biological activity upon storage. In one aspect, the formulation substantially retains its physical and chemical stability, as well as its biological activity upon storage. The storage period is generally selected based on the intended shelf-life of the formulation. Various analytical techniques for measuring protein stability are available in the art and are reviewed, for example, in Peptide and Protein Drug Delivery, 247-301, Vincent Lee Ed., Marcel Dekker, Inc., New York, N.Y., Pubs. (1991) and Jones, A. Adv. Drug Delivery Rev. 10:29-90 (1993). Stability can be measured at a selected temperature for a selected time period. For example, a liquid formulation is stable at about 40° C. for at least about 3 days, 5 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks or 6 weeks. In one embodiment, a liquid formulation is stable at about 5° C. and/or 25° C. for at least about 1 month, at least about 3 months, at least about 6 months, at least about 9 months, at least about 12 months, at least about 18 months, at least about 24 months, at least about 30 months, or at least about 36 months; and/or stable at about −20° C. and/or −70° C. for at least about 1 month, at least about 3 months, at least about 6 months, at least about 9 months, at least about 12 months, at least about 18 months, at least about 24 months, at least about 30 months, at least about 36 months, at least about 42 months, or at least about 48 months. Furthermore, the liquid formulation may, in some embodiments, be stable following freezing (to, e.g., −80° C.) and thawing, for example following 1, 2 or 3 cycles of freezing and thawing.

The stability of a liquid formulation can be evaluated qualitatively and/or quantitatively in a variety of different ways, including evaluation of dimer, multimer and/or aggregate formation (for example using size exclusion chromatography (SEC), matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS), analytical ultracentrifugation, light scattering (photon correlation spectroscopy, dynamic light scattering (DLS), static light scattering, multi-angle laser light scattering (MALLS)), flow-based microscopic imaging, electronic impedance (coulter) counting, light obscuration or other liquid particle counting system, by measuring turbidity, and/or by visual inspection); by assessing charge heterogeneity using cation exchange chromatography (CEX), isoelectric focusing (IEF), e.g. capillary technique (cIEF), or capillary zone electrophoresis; amino-terminal or carboxy-terminal sequence analysis; mass spectrometric analysis; SDS-PAGE or SEC analysis to compare fragmented, intact and multimeric (i.e., dimeric, trimeric, etc.) antibody; peptide map (for example tryptic or LYS-C) analysis; evaluating biological activity or antigen binding function of the antibody; and the like. Instability may involve any one or more of: aggregation, deamidation (e.g. Asn deamidation), oxidation (e.g. Met oxidation), isomerization (e.g. Asp isomeriation), clipping/hydrolysis/fragmentation (e.g. hinge region fragmentation), succinimide formation, N-terminal extension, C-terminal processing, glycosylation differences, and the like.

A “deamidated” monoclonal antibody is one in which one or more asparagine or glutamine residue thereof has been derivatized, e.g. to an aspartic acid or an iso-aspartic acid.

An antibody which is “susceptible to deamidation” is one comprising one or more residue which has been found to be prone to deamidate.

An antibody which is “susceptible to oxidation” is an antibody comprising one or more residue which has been found to be prone to oxidation.

An antibody which is “susceptible to aggregation” is one which has been found to aggregate with other antibody molecule(s), especially upon freezing, heating, drying, reconstituting and/or agitation.

An antibody which is “susceptible to fragmentation” is one which has been found to be cleaved into two or more fragments, for example at a hinge region thereof.

By “reducing deamidation” is intended to mean preventing or decreasing (e.g., to 80%, 60%, 50%, 40%, 30%, 20% or 10% of) the amount of deamidation relative to the monoclonal antibody formulated at a different pH or in a different buffer.

By “reducing oxidation” is intended to mean preventing or decreasing (e.g., to 80%, 60%, 50%, 40%, 30%, 20% or 10% of) the amount of oxidation relative to the monoclonal antibody formulated at a different pH or in a different buffer.

By “reducing aggregation” is intended to mean preventing or decreasing (e.g., to 80%, 60%, 50%, 40%, 30%, 20% or 10% of) the amount of aggregation relative to the monoclonal antibody formulated at a different pH or in a different buffer.

By “reducing fragmentation” is intended to mean preventing or decreasing (e.g., to 80%, 60%, 50%, 40%, 30%, 20% or 10% of) the amount of fragmentation relative to the monoclonal antibody formulated at a different pH or in a different buffer.

As used herein, the terms “aggregate” or “aggregates” refer to the association of two or more antibodies or antibody fragments. For example, an aggregate can be a dimer, trimer, tetramer, or a multimer greater than a tetramer, of antibodies and/or antibody fragments. Antibody aggregates can be soluble or insoluble. The association between the aggregated molecules may be either covalent or non-covalent without respect to the mechanism by which they are associated. The association may be direct between the aggregated molecules or indirect through other molecules that link them together. Examples of the latter include, but are not limited to disulfide linkages with other proteins, hydrophobic associations with lipids, charge associations with DNA, affinity associations with leached protein A, or mixed mode associations with multiple components. Aggregates can be irreversibly formed either during protein expression in cell culture, during protein purification in downstream processing, or during storage of the drug product. The presence of aggregates in a solution can be determined using, for example, size exclusion chromatography (SEC) (e.g., SEC with UV detection, SEC with light scattering detection (SEC-LSD)), field flow fractionation, analytical ultracentrifugation sedimentation velocity, or capillary electrophoresis-sodium dodecyl sulfate (CE-SDS, reduced and non-reduced).

The term “high molecular weight” or “HMW” is used to indicate an antibody complex having a molecular weight greater than a monomer antibody. In one embodiment, a HMW aggregate has a molecular weight greater than about 147 kDa. The presence of high molecular weight aggregates may be determined by standard methods known in the art, e.g., size-exclusion chromatography (SEC).

As used herein, “biological activity” of an antibody refers to the ability of the antibody to bind to antigen and result in a measurable biological response which can be measured in vitro or in vivo.

The cell surface molecule, “α4β7 integrin.” “alpha4beta7”, or “α4β7” (used interchangeably throughout) is a heterodimer of an α4 chain (CD49D, ITGA4) and a β7 chain (ITGB7). Human α4-integrin and β7-integrin genes GenBank (National Center for Biotechnology Information, Bethesda, Md.) RefSeq Accession numbers NM_000885 and NM_000889, respectively) are expressed by B and T lymphocytes, particularly memory CD4+ lymphocytes. Typical of many integrins, α4β7 can exist in either a resting or activated state. Ligands for α4β7 include vascular cell adhesion molecule (VCAM), fibronectin and mucosal addressin (MAdCAM (e.g., MAdCAM-1)). An antibody that binds to α4β7 integrin is referred to herein as an “anti-α4β7 antibody”.

As used herein, an antibody, or antigen-binding fragment thereof, that has “binding specificity for the α4β7 complex” binds to α4β7, but not specifically to α4β1 or α4β7. Vedolizumab is an example of an antibody that has binding specificity for the α4β7 complex.

As used herein, an “isotonic” formulation has substantially the same osmotic pressure as human blood. Isotonic formulations will generally have an osmotic pressure from about 250 to 350 mOsm. Isotonicity can be measured using a vapor pressure or ice-freezing type osmometer, for example.

As used herein, “viscosity” is a fluid's resistance to flow, and may be measured in units of centipoise (cP) or milliPascal-second (mPa-s), where 1 cP=1 mPa-s, at a given shear rate. Viscosity may be measured, for example, by using a viscometer. Viscosity may be measured using any other methods and in any other units known in the art (e.g. absolute, kinematic or dynamic viscosity), understanding that it is the percent reduction in viscosity afforded by use of the excipients described by the invention that is important. Regardless of the method used to determine viscosity, the percent reduction in viscosity in excipient formulations versus control formulations will remain approximately the same at a given shear rate.

As used herein, a liquid pharmaceutical formulation containing an amount of an excipient effective to “reduce viscosity” (or a “viscosity-reducing” amount or concentration of such excipient) means that the viscosity of the formulation in its final form for administration (e.g., a liquid solution) is at least 5% less than the viscosity of an appropriate control formulation, such as water, buffer, other viscosity-reducing agents such as salt, etc. and those control formulations, for example, exemplified herein. Similarly, a “reduced viscosity” formulation is a formulation that exhibits reduced viscosity compared to a control formulation.

As used herein, “buffering agent” refers to an agent that resists changes in pH by the action of its acid-base conjugate components. Examples of buffering agents include, but are not limited to, acetate, succinate, succinate, gluconate, histidine, citrate, glycylglycine, potassium phosphate, magnesium sulfate, carbonate, borate, phthalate, glutamate, and other organic acid buffers. In some embodiments, the buffering agent used in the formulations herein adjusts the pH of the formulation to about 5.0 to about 7.5, to about pH 5.5 to about 7.5, to about pH 6.0 to about 7.0, or to a pH of about 6.3 to about 6.5. In one aspect, examples of buffering agents that alone or in combination, will control the pH in the 5.0 to 7.5 range include acetate, succinate, gluconate, histidine, citrate, phosphate, maleate, cacodylate, 2-[N-morpholino]ethanesulfonic acid (MES), bis(2-hydroxyethyl)iminotris[hydroxymethyl]methane (Bis-Tris), N-[2-acetamido]-2-iminodiacetic acid (ADA), glycylglycine and other organic acid buffers. In one embodiment, a buffering agent does not include citrate. In one embodiment, a buffering agent does not include histidine.

A “histidine buffer” is a buffer comprising histidine ions. Examples of histidine buffers include histidine chloride, histidine acetate, histidine phosphate, histidine sulfate solutions. The histidine buffer or histidine-HCl buffer has a pH between about pH 5.5 to about 7.0, between about pH 6.1 to about 6.9, or about pH 6.5.

Herein, a “surfactant” refers to an agent that lowers surface tension of a liquid. In one aspect, the surfactant is a nonionic surfactant. Examples of surfactants herein include polysorbate (polyoxyethylene sorbitan monolaurate, for example, polysorbate 20 and polysorbate 80); TRITON (t-Octylphenoxypolyethoxyethanol, nonionic detergent, Union Carbide subsidiary of Dow Chemical Co., Midland Mich.); sodium dodecyl sulfate (SDS); sodium laurel sulfate; sodium octyl glycoside; lauryl-, myristyl-, linoleyl-, or stearyl-sulfobetaine; lauryl-, myristyl-, linoleyl- or stearyl-sarcosine; linoleyl-, myristyl-, or cetyl-betaine; lauroamidopropyl-, cocamidopropyl-, linolcamidopropyl-, myristamidopropyl-, palmidopropyl-, or isostearamidopropyl-betaine (e.g. lauroamidopropyl); myristamidopropyl-, palmidopropyl-, or isostearamidopropyl-dimethylamine; sodium methyl cocoyl-, or disodium methyl oleyl-taurate; sorbitan monopalmitate; and the MONAQUAT series (Mona Industries, Inc., Paterson, N.J.); polyethyl glycol (PEG), polypropylene glycol (PPG), and copolymers of poloxyethylene and poloxypropylene glycol (e.g. Pluronics/Poloxamer, PF68 etc); etc. In another aspect, the surfactant herein is polysorbate 80.

The term “chelator” refers to an agent that binds to an atom through more than one bond. In one aspect, examples of chelators herein include citrate, ethylenediaminetetraacetic acid, ethyleneglycoltetraacetic acid (EGTA), dimercaprol, diethylenetriaminepentaacetic acid, and N,N-bis(carboxymethyl)glycine. In another aspect, the chelator is not citrate.

The term “antioxidant” refers to an agent that inhibits the oxidation of other molecules. Examples of antioxidants herein include citrate, lipoic acid, uric acid, glutathione, tocopherol, carotene, lycopene, cysteine, phosphonate compounds, e.g., etidronic acid, desferoxamine and malate.

The term “antibody” as used herein, is intended to refer to an immunoglobulin molecule comprised of four polypeptide chains, two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds. Each heavy chain is comprised of a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region (CH). The heavy chain constant region is comprised of three domains, CH1, CH2 and CH3. Each light chain is comprised of a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region is comprised of one domain, CL. The VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDRs), interspersed with regions that are more conserved, termed framework regions (FR). Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In some embodiments, the antibody has a fragment crystallizable (Fc) region. In certain embodiments, the antibody is an IgG1 isotype and has a kappa light chain.

A “CDR” or “complementarity determining region” is a region of hypervariability interspersed within regions that are more conserved, termed “framework regions” (FR). Generally CDRs impart antibody binding specificity to a target antigen.

As used herein, the term “antigen binding fragment” or “antigen binding portion” of an antibody refers to Fab, Fab′, F(ab′), and Fv fragments, single chain antibodies, functional heavy chain antibodies (nanobodies), as well as any portion of an antibody having specificity toward at least one desired epitope, that competes with the intact antibody for specific binding (e.g., an isolated portion of a complementarity determining region having sufficient framework sequences so as to bind specifically to an epitope). Antigen binding fragments can be produced by recombinant techniques, or by enzymatic or chemical cleavage of an antibody.

“Humanized” forms of non-human (e.g., rodent) antibodies are chimeric antibodies that contain minimal sequence derived from the non-human antibody. For the most part, humanized antibodies are human immunoglobulins (recipient antibody) in which residues from a hypervariable region of the recipient are replaced by residues from a hypervariable region of a non-human species (donor antibody) such as mouse, rat, rabbit or nonhuman primate having the desired specificity, affinity, and capacity. In some instances, framework region (FR) residues of the human antibody are replaced by corresponding non-human residues. Furthermore, humanized antibodies may comprise residues that are not found in the recipient antibody or in the donor antibody. These modifications are made to further refine antibody performance. In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable CDR loops correspond to those of a non-human antibody and all or substantially all of the FRs are those of a human antibody sequence. The humanized antibody optionally also will comprise at least a portion of an antibody constant region (Fc), typically that of a human antibody. For further details, see Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992).